Figure 4. Translational regulation of PP1α cs and eIF6 in response to HCMV regulates productive viral growth.

(A) NHDFs were mock-infected or infected (MOI=3) with WT HCMV, a UL38-deficient mutant virus (deltaUL38), or a revertant virus where the UL38-deficiency was repaired by reintroducing a WT UL38 gene (UL38rev). At 48 hpi, total protein was isolated, fractionated by SDS-PAGE and analyzed by immunoblotting with the indicated antisera. Samples (n=3) were quantified using an Odyssey infrared imager. Each band was measured for raw intensity value, normalized to the loading control (Akt) and fold-change upon infection determined. (B) As in A except NHDFs were infected with WT HCMV and total protein harvested at the indicated HPI. (C) NHDFs transfected with a control, non-silencing siRNA (ctrl) or individual siRNAs targeting PP1α (#5, #6) were infected with HCMV (MOI=0.1). After 96 hpi samples were analyzed by immunoblotting as in A. D) Supernatants collected from three independent experiments detailed in C were assayed for viral particle production based upon their TCID50 (Kudchodkar et al., 2004). E) As in B. F) As in C but using siRNAs targeting PABP1 or eIF6. G) Supernatants collected from three independent experiments detailed in F were assayed for viral particle production as in D.